CN110527000A - A kind of method that polygonatum cyrtonema aerial part extracts polysaccharide - Google Patents
A kind of method that polygonatum cyrtonema aerial part extracts polysaccharide Download PDFInfo
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- CN110527000A CN110527000A CN201910863027.6A CN201910863027A CN110527000A CN 110527000 A CN110527000 A CN 110527000A CN 201910863027 A CN201910863027 A CN 201910863027A CN 110527000 A CN110527000 A CN 110527000A
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- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0003—General processes for their isolation or fractionation, e.g. purification or extraction from biomass
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Abstract
The invention discloses a kind of methods that polygonatum cyrtonema aerial part extracts polysaccharide, collect the aerial part of polygonatum cyrtonema, and the polygonatum cyrtonema raw material after drying crushes；Petroleum ether is added into polygonatum cyrtonema powder, shakes up rear water-bath reflux, filters recycling petroleum ether, after ethanol solution refluxing extraction is added into filter cake, collected by suction filter residue is dry；Polygonatum cyrtonema aerial part filter residue is uniformly mixed with distilled water, filters and collects filtrate；It is slowly added ethyl alcohol into smoke filtrate and constantly whisks, abandons supernatant, collects precipitating, precipitating sequentially adds ethyl alcohol, acetone, ether washing repeatedly, volatilizes；Polysaccharide precipitation is dissolved in distilled water, takes off albumen repeatedly, gained powder is polygonatum cyrtonema refined polysaccharide.The present invention can increase the output of polygonatum cyrtonema polysaccharide 40% compared to single polygonatum cyrtonema rhizome Polyose extraction, polygonatum cyrtonema aerial part.Polygonatum cyrtonema resource can be substantially more utilized using the present invention；The supply channel of polygonatum cyrtonema polysaccharide has also been widened simultaneously.
Description
Technical field
The present invention relates to a kind of extraction processes of polygonatum cyrtonema more particularly to a kind of polygonatum cyrtonema aerial part to extract
The method of polysaccharide.
Background technique
Polygonatum cyrtonema Polygonatumcyrtonema system Liliaceae HUANGJING ZANYU CAPSULE, with rhizoma polygonati P.sibiricum and Yunnan
Rhizoma polygonati P.kingianum is embodied in version " Chinese Pharmacopoeia " in 2015 together.In history, rhizoma polygonati is the perennial herb of integration of drinking and medicinal herbs
Plant.Its is sweet in flavor, flat, and returns spleen, lung, kidney channel have the benefits of boosting qi and nourishing yin, invigorating the spleen, moistening lung, kidney-nourishing.Modern research shows that rhizoma polygonati
In the ingredients such as polysaccharide rich in, flavones, saponin(e, volatile oil, main component polysaccharide has hypoglycemic and blood lipid, antibacterial,
It is anti-inflammatory, it is antitumor, enhance the effects of immune and antiviral.
It currently to the extracting method of polygonatum cyrtonema polysaccharide, is used after polygonatum cyrtonema medicinal material is sliced and is crushed
The link that solvent appropriate is extracted, refines and dried obtains Siberian solomonseal rhizome polysaccharide.These methods because use polygonatum cyrtonema medicinal material,
Production cost is relatively high.And the polygonatum cyrtonema aerial part equally containing abundant polysaccharide component can also be used as polygonatum cyrtonema
The important sources of polysaccharide.
Summary of the invention
Technical problem to be solved by the present invention lies in: it is not mentioned in the prior art using the aerial part of polygonatum cyrtonema
Polysaccharide is taken, a kind of method that polygonatum cyrtonema aerial part extracts polysaccharide is provided.
The present invention be by the following technical programs solution above-mentioned technical problem, the present invention the following steps are included:
(1) dry
The aerial part for collecting polygonatum cyrtonema is no more than 10% in 45~60 DEG C of dryings to water content；
(2) it crushes
By the polygonatum cyrtonema raw material after drying, 65 meshes are smashed it through；
(3) degreasing cleans
The petroleum ether of 10 times of quality is added into polygonatum cyrtonema powder, shakes up rear water-bath reflux, filters recycling petroleum ether, together
When volatilize unrecovered oil ether in filter cake, into filter cake be added 10 times of quality volume fraction be 95% ethanol solution refluxing extraction
Afterwards, collected by suction filter residue, 45~60 DEG C of dryings；
(4) it extracts
Polygonatum cyrtonema aerial part filter residue is uniformly mixed with the distilled water of 15 times of quality, refluxing extraction is multiple, filters simultaneously
Collect filtrate；
(5) concentrating and precipitating
Gained filtrate is concentrated under reduced pressure into 1/6 volume of initial filtrate under the conditions of 55~65 DEG C, slowly adds into smoke filtrate
Add 95% ethyl alcohol and constantly whisk, until ethyl alcohol final concentration of 80%, is centrifuged after 4 DEG C of placement 12h；
(6) it decolourizes
Supernatant is abandoned, precipitating is collected, precipitating sequentially adds ethyl alcohol, acetone, ether washing repeatedly, volatilizes；
(7) albumen and drying are taken off
Polysaccharide precipitation is dissolved in distilled water, takes off albumen repeatedly with Sevag reagent, 1/3 volume polysaccharide solution is added
Sevag reagent, it is repeated multiple times, until water phase and Sevag reagent liquid level are without white " milky " object, aqueous phase solution freeze-drying, gained
Powder is polygonatum cyrtonema refined polysaccharide.
The polygonatum cyrtonema aerial part is that acquisition in any year obtains in its growth period.Polygonatum cyrtonema medicinal material is often after planting
3-4 acquires rhizome；And polygonatum cyrtonema aerial part Polyose extraction, it can all be acquired and add in each year in its growth period
Work can increase benefit 50% compared to single rhizome Polyose extraction.
As one of preferred embodiment of the invention, in the step (3), bath temperature is 70~80 DEG C, and flow back 2~3h.
As one of preferred embodiment of the invention, in the step (4), refluxing extraction 2~3 times, 2~3h every time.
As one of preferred embodiment of the invention, in the step (7), chloroform in Sevag reagent: n-butanol=4:1.
The present invention has the advantage that the present invention by mentioning to polygonatum cyrtonema aerial part polysaccharide compared with prior art
It takes, polygonatum cyrtonema resource can be made full use of；Compared to single polygonatum cyrtonema rhizome Polyose extraction, polygonatum cyrtonema aerial part
The output of polygonatum cyrtonema polysaccharide 40% can be increased.By selecting optimal processing technology high efficiency extraction polygonatum cyrtonema aerial part
Polysaccharide can make full use of polygonatum cyrtonema resource；Simultaneously as having widened the supply channel of polygonatum cyrtonema polysaccharide, can increase
The market supply, and reduce polygonatum cyrtonema for commodity price, allow more consumer benefits.
Detailed description of the invention
Fig. 1 is glucose standard curve figure.
Specific embodiment
It elaborates below to the embodiment of the present invention, the present embodiment carries out under the premise of the technical scheme of the present invention
Implement, the detailed implementation method and specific operation process are given, but protection scope of the present invention is not limited to following implementation
Example.
Measure glucose standard curve.
DEXTROSE ANHYDROUS reference substance 33mg (dry to constant weight through 105 DEG C) accurately weighed first, then set in 100mL measuring bottle,
Add ultrapure water to dissolve and be diluted to scale, shakes up to get (containing DEXTROSE ANHYDROUS 0.33mg) in every 1mL in solution.Standard curve
Preparation: precision is measured to muddy product solution 0.1mL, 0.2mL, 0.3mL, 0.4mL, 0.5mL, 0.6mL, is respectively placed in 10mL tool plug
In scale test tube, 2.0mL is respectively added water to, is shaken up, 0.2% Anthrone Sulphuric acid solution is slowly added dropwise in ice-water bath to scale, mixes
It is even, it lets cool in postposition water-bath and keeps the temperature 10 minutes, take out, place 10 minutes cooling in ice-water bath in beaker, taking-up, with corresponding immediately
Reagent is blank control.According to UV-VIS spectrophotometry, absorbance is measured at 582nm wavelength.It is vertical sit with absorbance
Mark, sample concentration is abscissa, draw standard curve Fig. 1 to get.
Embodiment 1
The present embodiment is used to extract the polysaccharide of polygonatum cyrtonema aerial part, and detailed process is as follows:
(1) dry: to collect polygonatum cyrtonema aerial part, 55 DEG C of dryings to water content is no more than 10% in baking oven.
(2) crush: the polygonatum cyrtonema raw material after taking drying crushed No. four sieves.
(3) degreasing and removal of impurities: 10 times of amount petroleum ethers (60~90 DEG C of boiling range) are added into polygonatum cyrtonema powder, 75 after shaking up
DEG C water-bath flows back 2h, filters to recycle petroleum ether, while volatilizing unrecovered oil ether in filter cake.10 times of amounts are added into filter cake
After 95% ethanol solution refluxing extraction 1.5h, collected by suction filter residue is dry in 55 DEG C of baking ovens.
(4) it extracts: polygonatum cyrtonema aerial part filter residue being uniformly mixed with 15 times of amount distilled water, refluxing extraction 2 times, every time
2h is filtered and is collected filtrate.
(5) concentrating and precipitating: gained filtrate is concentrated under reduced pressure into 1/6 volume of initial filtrate under the conditions of 60 DEG C, to smoke filtrate
In be slowly added 95% ethyl alcohol and constantly whisk, until ethyl alcohol final concentration of 80%, be centrifuged after 4 DEG C of placement 12h.
(6) it decolourizes: abandoning supernatant, collect precipitating, precipitating sequentially adds ethyl alcohol, acetone, ether (20,10,10mL) repeatedly and washes
It washs three times, volatilizes.
(7) take off albumen and drying: polysaccharide precipitation is re-dissolved in appropriate distilled water (30mL), with Sevag reagent (chloroform: just
Butanol=4:1) repeatedly take off albumen, be added 1/3 volume polysaccharide solution Sevag reagent, 3~5 times repeatedly, until water phase and Sevag
Reagent liquid level is without white " milky " object, and aqueous phase solution freeze-drying, gained powder is polygonatum cyrtonema refined polysaccharide.
Embodiment 2
The present embodiment is used to extract the polysaccharide of polygonatum cyrtonema aerial part, and detailed process is as follows:
(1) dry: to collect polygonatum cyrtonema aerial part, 55 DEG C of dryings to water content is no more than 10% in baking oven.
(2) crush: the polygonatum cyrtonema raw material after taking drying crushed No. four sieves.
(3) degreasing and removal of impurities: 10 times of amount petroleum ethers (60~90 DEG C of boiling range) are added into polygonatum cyrtonema powder, 75 after shaking up
DEG C water-bath flows back 2h, filters to recycle petroleum ether, while volatilizing unrecovered oil ether in filter cake.10 times of amounts are added into filter cake
After 95% ethanol solution refluxing extraction 1.5h, collected by suction filter residue is dry in 55 DEG C of baking ovens.
(4) it extracts: polygonatum cyrtonema aerial part filter residue being uniformly mixed with 15 times of amount distilled water, refluxing extraction 2 times, every time
2h is filtered and is collected filtrate.
(5) concentrating and precipitating: gained filtrate is concentrated under reduced pressure into 1/6 volume of initial filtrate under the conditions of 60 DEG C, to smoke filtrate
In be slowly added 95% ethyl alcohol and constantly whisk, until ethyl alcohol final concentration of 80%, be centrifuged after 4 DEG C of placement 12h.
(6) take off albumen and drying: polysaccharide precipitation is re-dissolved in appropriate distilled water (30mL), with Sevag reagent (chloroform: just
Butanol=4:1) repeatedly take off albumen, be added 1/3 volume polysaccharide solution Sevag reagent, 3~5 times repeatedly, until water phase and Sevag
Reagent liquid level is without white " milky " object, and aqueous phase solution freeze-drying, gained powder is polygonatum cyrtonema refined polysaccharide.
Embodiment 3
The present embodiment is used to extract the polysaccharide of polygonatum cyrtonema aerial part, and detailed process is as follows:
(1) dry: to collect polygonatum cyrtonema aerial part, 55 DEG C of dryings to water content is no more than 10% in baking oven.
(2) crush: the polygonatum cyrtonema raw material after taking drying crushed No. four sieves.
(3) it extracts: polygonatum cyrtonema aerial part filter residue being uniformly mixed with 15 times of amount distilled water, refluxing extraction 2 times, every time
2h is filtered and is collected filtrate.
(4) concentrating and precipitating: gained filtrate is concentrated under reduced pressure into 1/6 volume of initial filtrate under the conditions of 60 DEG C, to smoke filtrate
In be slowly added 95% ethyl alcohol and constantly whisk, until ethyl alcohol final concentration of 80%, be centrifuged after 4 DEG C of placement 12h.
(5) it decolourizes: abandoning supernatant, collect precipitating, precipitating sequentially adds ethyl alcohol, acetone, ether (20,10,10mL) repeatedly and washes
It washs three times, volatilizes.
(6) take off albumen and drying: polysaccharide precipitation is re-dissolved in appropriate distilled water (30mL), with Sevag reagent (chloroform: just
Butanol=4:1) repeatedly take off albumen, be added 1/3 volume polysaccharide solution Sevag reagent, 3~5 times repeatedly, until water phase and Sevag
Reagent liquid level is without white " milky " object, and aqueous phase solution freeze-drying, gained powder is polygonatum cyrtonema refined polysaccharide.
Embodiment 4
The present embodiment is used to extract the polysaccharide of polygonatum cyrtonema aerial part, and detailed process is as follows:
(1) dry: to collect polygonatum cyrtonema aerial part, 55 DEG C of dryings to water content is no more than 10% in baking oven.
(2) crush: the polygonatum cyrtonema raw material after taking drying crushed No. four sieves.
(3) degreasing and removal of impurities: 10 times of amount petroleum ethers (60~90 DEG C of boiling range) are added into polygonatum cyrtonema powder, 75 after shaking up
DEG C water-bath flows back 2h, filters to recycle petroleum ether, while volatilizing unrecovered oil ether in filter cake.10 times of amounts are added into filter cake
After 95% ethanol solution refluxing extraction 1.5h, collected by suction filter residue is dry in 55 DEG C of baking ovens.
(4) it extracts: polygonatum cyrtonema aerial part filter residue being uniformly mixed with 15 times of amount distilled water, refluxing extraction 2 times, every time
2h is filtered and is collected filtrate.
(5) concentrating and precipitating: gained filtrate is concentrated under reduced pressure into 1/6 volume of initial filtrate under the conditions of 60 DEG C, to smoke filtrate
In be slowly added 95% ethyl alcohol and constantly whisk, until ethyl alcohol final concentration of 80%, be centrifuged after 4 DEG C of placement 12h.
(6) it decolourizes: abandoning supernatant, collect precipitating, precipitating sequentially adds ethyl alcohol, acetone, ether (20,10,10mL) repeatedly and washes
It washs three times, volatilizes.
Gained powder is polygonatum cyrtonema refined polysaccharide.
From Examples 1 to 4 as can be seen that embodiment 1 contains decoloration degreasing and Deproteinated all processing, embodiment 2
Decolorization is not carried out, and embodiment 3 does not carry out ungrease treatment, and embodiment 4 does not carry out de- albumen processing.
By the absorbance of measurement test sample, the content that Thick many candies and refined polysaccharide are calculated in standard curve is substituted into,
The results are shown in Table 1, from table it follows that embodiment 2 compared with Example 1, Thick many candies and refined polysaccharide content difference are little,
But the refined polysaccharide color that embodiment 2 (no bleaching) obtains is deeper；Embodiment 3 compared with Example 1, embodiment 3 (non-degreasing)
Middle Thick many candies content significantly increases, and illustrates have the substances such as other lipids to be also extracted；Embodiment 4 is compared with Example 1, real
Applying refined polysaccharide content in example 4 (not taking off albumen) significantly reduces, and illustrates to lead to polyoses content containing substances such as albumen in extracting solution
It reduces.To further demonstrate decoloration, degreasing, take off albumen for the necessity of the extraction of polygonatum cyrtonema aerial part polysaccharide.
Influence of 1 single factor test of table to polyoses content
Note: n=3, p < 0.05
It calculates: conversion factor (F): F=W/ (C × D).
Wherein: W: refined polysaccharide quality (mg)；C: glucose converts into concentration (mg/ml) in polysaccharide liquid；D: extension rate.It surveys
Glucose is obtained to the conversion factor of test sample polysaccharide.Sample is more to test sample multiplied by glucose with glucose meter polyoses content
The conversion factor of sugar, calculates polyoses content in test sample.
The foregoing is merely illustrative of the preferred embodiments of the present invention, is not intended to limit the invention, all in essence of the invention
Made any modifications, equivalent replacements, and improvements etc., should all be included in the protection scope of the present invention within mind and principle.
Claims (5)
1. a kind of method that polygonatum cyrtonema aerial part extracts polysaccharide, which comprises the following steps:
(1) dry
The aerial part for collecting polygonatum cyrtonema is no more than 10% in 45~60 DEG C of dryings to water content；
(2) it crushes
By the polygonatum cyrtonema raw material after drying, 65 meshes are smashed it through；
(3) degreasing cleans
The petroleum ether of 10 times of quality is added into polygonatum cyrtonema powder, shakes up rear water-bath reflux, filters recycling petroleum ether, wave simultaneously
Unrecovered oil ether in dry filter cake, the volume fraction that 10 times of quality are added into filter cake is pumping after 95% ethanol solution refluxing extraction
Filter residue, 45~60 DEG C of dryings are collected in filter；
(4) it extracts
Polygonatum cyrtonema aerial part filter residue is uniformly mixed with the distilled water of 15 times of quality, refluxing extraction is multiple, filters and collects
Filtrate；
(5) concentrating and precipitating
Gained filtrate is concentrated under reduced pressure into 1/6 volume of initial filtrate under the conditions of 55~65 DEG C, is slowly added into smoke filtrate
95% ethyl alcohol simultaneously constantly whisks, until ethyl alcohol final concentration of 80%, is centrifuged after 4 DEG C of placement 12h；
(6) it decolourizes
Supernatant is abandoned, precipitating is collected, precipitating sequentially adds ethyl alcohol, acetone, ether washing repeatedly, volatilizes；
(7) albumen and drying are taken off
Polysaccharide precipitation is dissolved in distilled water, takes off albumen repeatedly with Sevag reagent, and the Sevag examination of 1/3 volume polysaccharide solution is added
Agent, repeated multiple times, until water phase and Sevag reagent liquid level are without white " milky " object, aqueous phase solution is freeze-dried, and gained powder is
Polygonatum cyrtonema refined polysaccharide.
2. the method that a kind of polygonatum cyrtonema aerial part according to claim 1 extracts polysaccharide, which is characterized in that described more
Flower rhizoma polygonati aerial part is that acquisition in any year obtains in its growth period.
3. the method that a kind of polygonatum cyrtonema aerial part according to claim 1 extracts polysaccharide, which is characterized in that the step
Suddenly in (3), bath temperature is 70~80 DEG C, and flow back 2~3h.
4. the method that a kind of polygonatum cyrtonema aerial part according to claim 1 extracts polysaccharide, which is characterized in that the step
Suddenly in (4), refluxing extraction 2~3 times, 2~3h every time.
5. the method that a kind of polygonatum cyrtonema aerial part according to claim 1 extracts polysaccharide, which is characterized in that the step
Suddenly in (7), chloroform in Sevag reagent: n-butanol=4:1.
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| CN111978427A (en) * | 2020-09-08 | 2020-11-24 | 上海铮信生物科技有限公司 | Extraction method of polygonatum polysaccharide |
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| CN113144089A (en) * | 2021-03-11 | 2021-07-23 | 安徽中医药大学 | Pharmaceutical composition for inhibiting obesity and preparation method and application thereof |
| CN113501890A (en) * | 2021-07-19 | 2021-10-15 | 上海应用技术大学 | Extraction method of polygonatum cyrtonema polysaccharide and application of polygonatum cyrtonema polysaccharide |
| CN113966834A (en) * | 2021-07-02 | 2022-01-25 | 厦门医学院 | Polygonatum cyrtonema processing or preprocessing method and application thereof |
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| CN116162180A (en) * | 2023-02-15 | 2023-05-26 | 李晓春 | Polysaccharide and method for extracting polysaccharide from selenium-enriched Polygonatum cyrtonema Fabricius |
| CN116284464A (en) * | 2021-12-13 | 2023-06-23 | 广西中医药大学 | Polygonatum cyrtonema polysaccharide and preparation method and application thereof |
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| CN117801132A (en) * | 2024-01-02 | 2024-04-02 | 安徽中医药大学 | Polygonatum cyrtonema leaf polysaccharide and preparation method and application thereof |
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| CN111978427A (en) * | 2020-09-08 | 2020-11-24 | 上海铮信生物科技有限公司 | Extraction method of polygonatum polysaccharide |
| CN112755044A (en) * | 2021-01-27 | 2021-05-07 | 安徽中医药大学 | Pharmaceutical composition for reducing blood sugar, preparation method and application |
| CN113144089A (en) * | 2021-03-11 | 2021-07-23 | 安徽中医药大学 | Pharmaceutical composition for inhibiting obesity and preparation method and application thereof |
| CN113966834B (en) * | 2021-07-02 | 2023-10-20 | 厦门医学院 | Polygonatum cyrtonema processing or preprocessing method and application thereof |
| CN113966834A (en) * | 2021-07-02 | 2022-01-25 | 厦门医学院 | Polygonatum cyrtonema processing or preprocessing method and application thereof |
| CN113501890B (en) * | 2021-07-19 | 2022-07-12 | 上海应用技术大学 | Extraction method of polygonatum cyrtonema polysaccharide and application of polygonatum cyrtonema polysaccharide |
| CN113501890A (en) * | 2021-07-19 | 2021-10-15 | 上海应用技术大学 | Extraction method of polygonatum cyrtonema polysaccharide and application of polygonatum cyrtonema polysaccharide |
| CN116284464A (en) * | 2021-12-13 | 2023-06-23 | 广西中医药大学 | Polygonatum cyrtonema polysaccharide and preparation method and application thereof |
| CN114703241A (en) * | 2021-12-31 | 2022-07-05 | 安徽中医药大学 | Preparation method and application of polygonatum cyrtonema polysaccharide processed with wine |
| CN114703241B (en) * | 2021-12-31 | 2024-05-10 | 安徽中医药大学 | Preparation method and application of wine-processed polygonatum sibiricum polysaccharide |
| CN116162180A (en) * | 2023-02-15 | 2023-05-26 | 李晓春 | Polysaccharide and method for extracting polysaccharide from selenium-enriched Polygonatum cyrtonema Fabricius |
| CN116284488A (en) * | 2023-05-05 | 2023-06-23 | 曲靖医学高等专科学校 | Extraction method of Polygonatum kingianum polysaccharide |
| CN117801132A (en) * | 2024-01-02 | 2024-04-02 | 安徽中医药大学 | Polygonatum cyrtonema leaf polysaccharide and preparation method and application thereof |
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